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目的对弓形虫人源性RH株、人源性ZS_2株、猪源性CN株3个不同地理株的GRA1基因片段鉴定并比较异同。方法采用PCR技术和限制性内切酶技术,分别对弓形虫3株的GRA1基因片段进行体外扩增。扩增的目的基因片段分别用3种内切酶HindⅢ、HPaⅡ、TaqⅠ进行单酶切鉴定、比较。结果从弓形虫3个分离株RH株、ZS_2株、CN标基因组DNA中扩增出785bp的GRA1基因片段。3个株的GRA1基因分别用3种内切酶酶切后,酶切片段与理论值相符。结论实验所用不同来源弓形虫分离株的GRA1基因扩增片段基本相同,且3种限制性内切酶酶切图诺基本一致,表明这3个分离株的GRA1基因高度保守。
Objective To identify and compare the GRA1 gene fragments of three geographical isolates of human RH strain, human ZS 2 strain and porcine CN strain. Methods PCR and restriction endonuclease techniques were used to amplify the GRA1 gene fragment of three Toxoplasma gondii strains in vitro. The amplified target gene fragment was identified by three restriction enzymes HindⅢ, HPaⅡ and TaqⅠ, respectively. Results The GRA1 gene fragment of 785bp was amplified from 3 isolates of Toxoplasma gondii RH strain, ZS 2 strain and CN genomic DNA. Three strains of GRA1 gene were digested with three kinds of endonucleases, digested fragments consistent with the theoretical value. Conclusion The amplified fragment of GRA1 gene from Toxoplasma gondii isolates from different origins was basically the same, and the restriction enzyme digests of the three restriction enzymes were basically identical, indicating that the GRA1 gene of the three isolates was highly conserved.